Intra-tissular profile of adrenal steroids reveals variable levels associated with adrenocortical tumors (ACT) differentiation and suggests alterations in steroids export

Fideline Bonnet-Serrano; Louis Thomeret; Nesrine Benanteur; Patricia Vaduva; Lucas Bouys; Bruno Ragazzon; Annabel Berthon; Karine Perlemoine; Corinne Zientek; Martin Gaillard; Florian Violon; Mathilde Sibony; Christelle Laguillier-Morizot; Marie-Claude Menet; Lionel Groussin; Jean Guibourdenche; Anne Jouinot; Guillaume Assie; Jerome Bertherat

doi:10.1530/endoabs.99.P19

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Endocrine Abstracts (2024) 99 P19 | DOI: 10.1530/endoabs.99.P19

ECE2024 Poster Presentations Adrenal and Cardiovascular Endocrinology (95 abstracts)

Intra-tissular profile of adrenal steroids reveals variable levels associated with adrenocortical tumors (ACT) differentiation and suggests alterations in steroids export

Fidéline Bonnet-Serrano ^1,2 , Louis Thomeret ¹ , Nesrine Benanteur ¹ , Patricia Vaduva ¹ , Lucas Bouys ^1,3 , Bruno Ragazzon ¹ , Annabel Berthon ¹ , Karine Perlemoine ¹ , Corinne Zientek ² , Martin Gaillard ^1,4 , Florian Violon ^1,5 , Mathilde Sibony ^1,5 , Christelle Laguillier-Morizot ² , Marie-Claude Menet ⁶ , Lionel Groussin ^1,3 , Jean Guibourdenche ² , Anne Jouinot ¹ , Guillaume Assié ^1,3 & Jérôme Bertherat ^1,3

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Author affiliations

¹Institut Cochin, inserm u 1016, cnrs umr8104, Université Paris Cité, Equipe Génomique et Signalisation des Tumeurs Endocrines, Paris, France; ²APHP-Cochin Hospital, Hormonology, Paris, France; ³APHP-Cochin Hospital, Endocrinology, Paris, France; ⁴APHP-Cochin Hospital, Visceral Surgery, Paris, France; ⁵APHP-Cochin Hospital, Anatomopathology, Paris, France; ⁶Institut de Chimie Physique, Université Paris-Saclay-CNRS, UMR8000, Orsay, France

Introduction: Adrenal steroidogenesis is alterated in adrenocortical tumors (ACT), leading frequently to abnormal steroid secretion. The development of steroid assays by mass spectrometry (MS) has greatly advanced the characterization of these alterations. However, steroid profiles are classically performed in blood and urine, providing a limited insight into adrenal steroidogenesis, altered by the mixing with gonadal steroids and by the steroid peripheral catabolism. The objective of our work was to evaluate the steroids tissue level in normal adrenals and ACT to better characterize adrenal steroidogenesis process dysregulation.

Materials and methods: Fifty-five samples of fresh-frozen adrenocortical tissue stored in liquid nitrogen in the COMETE tumor bank were included: 9 samples of normal adrenals, 22 samples from adrenocortical carcinomas (ACC) and 24 samples from adrenocortical adenomas (including 10 cortisol-producing adenomas (CPA) and 14 non/mild autonomous cortisol secreting adenomas (NFAT/MACS)). After tissue grinding in liquid nitrogen and sonication in methanol, tissue extracts were injected into a TSQ-ALTIS instrument to determine a profile of 13 steroids (cortisol, 11-deoxycortisol, 17-hydroxyprogesterone, cortisone, 21-deoxycortisol, androstenedione, testosterone, corticosterone, 11-deoxycorticosterone, progesterone, 11-hydroxyandrostenedione, 11-ketoandrostenedione, 11-ketotestosterone) in UPLC–MS/MS. Steroid intratissular concentrations were expressed in nmol/kg of tissue allowing the comparison with blood concentrations in nmol/l.

Results: In tissue of normal adrenals, cortisol was the most abundant steroid as in peripheral blood. However, the ranges of intratissular values observed (median [IQR] =18790 [5252–27987] nmol/kg) were 10 to 100 times higher than the usual value of cortisol blood concentration. The other most abundant steroids in normal adrenocortical tissue were corticosterone (6951 [1454–8840] nmol/kg) and 11OH-androstenedione (2189 [1323–9595] nmol/kg). The ratios of intratissular levels to usual blood concentrations were higher for precursors (progesterone, 17OH-progesterone and 11-deoxycortisol) than for cortisol, suggesting a facilitated export of bioactive products. In ACT, besides the high intratissular concentrations measured, the most striking observation was the lower steroid levels measured in ACC tissues in comparison to benign adenomas’ tissues. Thus, intratissular cortisol level (nmol/kg) was lower in ACC (2655 [692–7933]) than in benign adenomas: CPA (27066 [21982–64731], Benjamini, Krieger and Yekutieli q<0.0001), MACS/NFAT (34241 [20425–43727], q<0.0001), and normal adrenals (q=0.036).

Discussion: This first study of tissue steroid assay in ACT reveals quantitative and qualitative differences among the tumor types. These differences are likely to be explained by the various mechanisms of tumor cells differenciation. The striking differences in the ratio of intratissular to blood levels between bioactive steroids and their precursors suggest differences in the regulation of steroids export.